title: Integrated proteomic and metabolic analysis of breast cancer progression.
abstract: One of the most persistent hallmarks of cancer biology is the preference of tumor cells to derive energy through glycolysis as opposed to the more efficient process of oxidative phosphorylation (OXPHOS). However, little is known about the molecular cascades by which oncogenic pathways bring about this metabolic switch. We carried out a quantitative proteomic and metabolic analysis of the MCF10A derived cell line model of breast cancer progression that includes parental cells and derivatives representing three different tumor grades of Ras-driven cancer with a common genetic background. A SILAC (Stable Isotope Labeling by Amino acids in Cell culture) labeling strategy was used to quantify protein expression in conjunction with subcellular fractionation to measure dynamic subcellular localization in the nucleus, cytosol and mitochondria. Protein expression and localization across cell lines were compared to cellular metabolic rates as a measure of oxidative phosphorylation (OXPHOS), glycolysis and cellular ATP. Investigation of the metabolic capacity of the four cell lines revealed that cellular OXPHOS decreased with breast cancer progression independently of mitochondrial copy number or electron transport chain protein expression. Furthermore, glycolytic lactate secretion did not increase in accordance with cancer progression and decreasing OXPHOS capacity. However, the relative expression and subcellular enrichment of enzymes critical to lactate and pyruvate metabolism supported the observed extracellular acidification profiles. This analysis of metabolic dysfunction in cancer progression integrated with global protein expression and subcellular localization is a novel and useful technique for determining organelle-specific roles of proteins in disease.
authors.affiliation: University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, United States of America ; Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Patrick G
authors.initials: PG
authors.lastname: Shaw
authors.forename: Raghothama
authors.initials: R
authors.lastname: Chaerkady
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.forename: Shauna
authors.initials: S
authors.lastname: Vasilatos
authors.forename: Yi
authors.initials: Y
authors.lastname: Huang
authors.forename: Bennett
authors.initials: B
authors.lastname: Van Houten
authors.forename: Akhilesh
authors.initials: A
authors.lastname: Pandey
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Davidson
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism / Q000503:physiopathology*; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D018450:Disease Progression; D004798:Enzymes / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D007553:Isotope Labeling / Q000379:methods; D055432:Metabolomics / Q000379:methods*; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D040901:Proteomics / Q000379:methods*; D013347:Subcellular Fractions / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D004798:Enzymes; D033681:Mitochondrial Membrane Transport Proteins